Product details

Product details

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Purpose

Convenient real-time RNA quantification in one EASY step.

Specificity

- Streamlined protocol in a simple, single-tube reaction set-up- High-quality, full-length cDNA from as little as 0.01pg of RNA template- Fully optimized for detection of low-copy genes- Simple set-up for any RNA template (e.g. mRNA and total RNA) - Reduces pipetting steps to minimize the risk of contamination and error.

Characteristics

One-Step RT-PCR Kit contains all necessary reagents for both reverse transcription and PCR amplification in a single reaction tube. Specifically, the One-Step RT-PCR Kit combines EasyScriptTM RTase and BestaqTM DNA Polymerase in a convenient format for highly sensitive and specific RT-PCR with any RNA template. Our proprietary RT-PCR Buffer contains stabilizers and enhancers that optimize the two reactions into a "single step". Together with our specially formulated RT-PCR Buffer, this One-Step RT-PCR Kit offers endusers an efficient, easy to use, and reliable alternative to conventional "twostep" sequential RT-PCR.

Application Details

100 rxn (20 μl/rxn)1. Aliquot reagents to avoid risk of contamination and repeated freeze-thaw cycles.2. Avoid high primer concentrations, a concentration of 100 nM to 300 nM of each primer usually gives the best results.3. Probe concentration may require optimization, typically a 3:1 to 1:1 primer:probe ratio will give good amplification.4. A very effective way to get tight Ct among replicates is to reduce pipetting error, this can be achieved by: 1) preparing amplicon specific pre-mix, 2) using repeating pipettes and 3) keeping the pipetting volume within the manufacturer's suggested range.5. For optimal results, it is recommended that the primers are 18-22 nucleotides in length with a Tm of 58°C-60°C and the size of target is about 100-250 bp.